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National Repository of Grey Literature

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Overview of methodological approaches leading to qualitative anad quantitative microbial biofilm evaluation Holmanová, Pavlína ; Konečná, Klára (advisor) ; Vejsová, Marcela (referee) Charles Univerzity Faculty of Pharmacy in Hradec Králové Study program: Medical Laboratory Technician Author: Pavlína Holmanová Supervisor: RNDr. Klára Konečná, Ph.D. Title of bachelor thesis: Overview of methodological approaches leading to qualitative and quantitative microbial biofilm evaluation Background: The aim of this work was to compose an overview of methodical approaches leading to qualitative and quantitative evaluation of microbial biofilms. To optimize the methods leading to preparation of mounts with biofilm-forming microorganisms evaluated by epifluorescence or confocal laser scanning microscopy. Methods: Optimization of microbial biofilm staining procedures for subsequent evaluation by epifluorescence microscopy. Formation of microbial biofilms - cultivation of microorganisms in various cultivation media in a microtiter plate, preparation of solutions with fluorescent dyes. Observation of coloured, dispersed biofilms by fluorescence microscope. Results: Using fluorescence microscopy, optimal target concentrations of fluorescent dyes in various combinations were verified. The combination of Syto 9 (SYTO™ 9 Green Fluorescent Nucleic Acid Stain) and propidium iodide was confirmed to be effective for the determination of viability of biofilm-forming microbial cells. The combination of... Detailed record

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